Antiviral drugs for treatment or prevention of dengue infection

ABSTRACT

Compounds, methods and pharmaceutical compositions for treating viral infections, by administering certain compounds in therapeutically effective amounts are disclosed. Methods for preparing the compounds and methods of using the compounds and pharmaceutical compositions thereof are also disclosed. In particular, the treatment and prophylaxis of viral infections such as caused by flavivirus is disclosed, i.e., including but not limited to, Dengue virus, West Nile virus, yellow fever virus, Japanese encephalitis virus, and tick-borne encephalitis virus.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a divisional of U.S. application Ser. No. 12/601,187filed Apr. 5, 2010, which is a national stage filing of correspondinginternational application number PCT/US2008/064662, filed on May 23,2008, which claims priority of U.S. Provisional Application No.60/924,628, filed May 23, 2007, all of which are hereby incorporated byreference in their entirety.

FIELD OF THE INVENTION

This invention relates to the use of benzenesulfonamide derivatives andanalogs, as well as compositions containing the same, for the treatmentor prophylaxis of viral diseases associated with the flavivirus familysuch as Dengue fever, Yellow fever, West Nile, St. Louis encephalitis,Hepatitis C, Murray Valley encephalitis, and Japanese encephalitis.

BACKGROUND OF THE INVENTION

Dengue fever (DF) is an acute febrile disease caused by one of fourclosely related virus serotypes (DEN-1, DEN-2, DEN-3, and DEN-4). Denguefever is classified based on its clinical characteristics into classicaldengue fever, or the more severe forms, dengue hemorrhagic feversyndrome (DHF), and dengue shock syndrome (DSS). Recovery from infectionfrom one serotype produces life-long immunity to that particularserotype, but provides only short-lived and limited protection againstany of the other serotypes (37). Dengue is a member of the Flaviviridaefamily which are enveloped, positive-sense RNA viruses whose humanpathogens also include West Nile virus (WNV), yellow fever virus (YFV),Japanese encephalitis virus (JEV), and tick-borne encephalitis virus(TBEV) among others. Dengue transmission is via the bite of an infectedAedes aegypti mosquito which is found in tropical and sub-tropicalregions around the world.

Each year regional epidemics of dengue cause significant morbidity andmortality, social disruption and substantial economic burden on thesocieties affected both in terms of hospitalization and mosquitocontrol. Dengue is considered by the World Health Organization (WHO) tobe the most important arthropod-borne viral disease with an estimated 50million cases of dengue infection, including 500,000 DHF cases and24,000 deaths worldwide each year (37, 38). WHO estimates that fortypercent of the world's population (2.5 billion people) are at risk forDF, DHF, and DSS (37). Dengue is also a NIAID Category A pathogen and interms of bio-defense, represents a significant threat to United Statestroops overseas. Dengue is an emerging threat to North America with adramatic increase in severe disease in the past 25 years including majorepidemics in Cuba and Venezuela, and outbreaks in Texas and Hawaii (4).Failure to control the mosquito vector and increases in long-distancetravel have contributed to the increase and spread of dengue disease.The characteristics of dengue as a viral hemorrhagic fever virus(arthropod-borne, widely spread, and capable of inducing a great amountof cellular damage and eliciting an immune response that can result insevere hemorrhage, shock, and death) makes this virus a unique threat todeployed military personnel around the world as well as to travelers totropical regions. Preparedness for both biodefense and for the publichealth challenges posed by dengue will require the development of newvaccines and antiviral therapeutics.

Dengue causes several illnesses with increasing severity beingdetermined in part by prior infection with a different serotype of thevirus. Classic dengue fever (DF) begins 3-8 days after the bite of aninfected mosquito and is characterized by sudden onset of fever,headache, back pain, joint pain, a measles-like rash, and nausea andvomiting (21). DF is frequently referred to as “breakbone” fever due tothese symptoms. The disease usually resolves after two weeks but aprolonged recovery with weakness and depression is common. The moresevere form of the disease, dengue hemorrhagic fever (DHF) has a similaronset and early phase of illness as dengue fever. However, shortly afteronset the disease is characterized by high fever, enlargement of theliver, and hemorrhagic phenomena such as bleeding from the nose, mouth,and internal organs due to vascular permeability (38). In dengue shocksyndrome (DSS) circulatory failure and hypovolaemic shock resulting fromplasma leakage occur and can lead to death in 12-24 hours without plasmareplacement (38). The case fatality rate of DHF/DSS can be as high as20% without treatment. DHF has become a leading cause of hospitalizationand death among children in many countries with an estimated 500,000cases requiring hospitalization each year and a case fatality rate ofabout 5% (37).

The pathogenesis of DHF/DSS is still being studied but is thought to bedue in part to an enhancement of virus replication in macrophages byheterotypic antibodies, termed antibody-dependent enhancement (ADE) (8).During a secondary infection, with a different serotype of dengue virus,cross-reactive antibodies that are not neutralizing form virus-antibodycomplexes that are taken into monocytes and Langerhans cells (dendriticcells) and increase the number of infected cells (7). This leads to theactivation of cytotoxic lymphocytes which can result in plasma leakageand the hemorrhagic features characteristic of DHF and DSS (21). Thisantibody-dependent enhancement of infection is one reason why thedevelopment of a successful vaccine has proven to be so difficult.Although less frequent, DHF/DSS can occur after primary infection (33),so virus virulence (16) and immune activation are also believed tocontribute to the pathogenesis of the disease (26).

Dengue is endemic in more than 100 countries in Africa, the Americas,the Eastern Mediterranean, South-east Asia and the Western Pacific.During epidemics, attack rates can be as high as 80-90% of thesusceptible population. All four serotypes of the virus are emergingworldwide, increasing the number of cases of the disease as well as thenumber of explosive outbreaks. In 2002 for example, there were 1,015,420reported cases of dengue in the Americas alone with 14,374 cases of DHF,which is more than three times the number of dengue cases reported inthe Americas in 1995 (24).

The dengue genome, approximately 11 kb in length, consists of a linear,single stranded, infectious, positive sense RNA that is translated as asingle long polyprotein (reviewed in (29). The genome is composed ofseven nonstructural (NS) protein genes and three structural proteingenes which encode the nucleocapsid protein (C), a membrane-associatedprotein (M), and an envelope protein (E). The nonstructural proteins areinvolved in viral RNA replication (35), viral assembly, and theinflammatory components of the disease (19). The structural proteins areinvolved mainly in viral particle formation (22). The precursorpolyprotein is cleaved by cellular proteinases to separate thestructural proteins (18), while a virus-encoded proteinase cleaves thenonstructural region of the polyprotein (6). The genome is capped anddoes not have a poly(A) tail at the 3′ end but instead has a stablestem-loop structure necessary for stability and replication of thegenomic RNA (3). The virus binds to cellular receptors via the E proteinand undergoes receptor-mediated endocytosis followed by low-pH fusion inlysosomes (20). The viral genome is then uncoated and translated intothe viral precursor polyprotein. Co- and posttranslational proteolyticprocessing separates the structural and nonstructural proteins. TheRNA-dependent RNA polymerase along with cofactors synthesizes theminus-strand RNA which serves as a template for the synthesis of theprogeny plus-strand RNA (25). Viral replication is membrane associated(1, 34). Following replication, the genome is encapsidated, and theimmature virus, surrounded by a lipid envelope buds into the lumen (9).The envelope proteins become glycosylated and mature viruses arereleased outside the cell. Essential stages or process during the viruslife cycle would be possible targets for inhibition from an antiviraldrug and include binding of the virus to the cell through the E protein,uptake of the virus into the cell, the capping mechanism, the viralproteinase, the viral RNA-dependent RNA polymerase, and the viralhelicase.

Current management of dengue virus-related disease relies solely onvector control. There are no approved antivirals or vaccines for thetreatment or prevention of dengue. Ribavirin, a guanosine analogue, hasbeen shown to be effective against a range of RNA virus infections andworks against dengue in tissue culture by inhibiting the dengue2′-O-methyltransferase NS5 domain (2, 10). However, ribavirin did notshow protection against dengue in a mouse model (15) or a rhesus monkeymodel (17), instead it induced anemia and thrombocytosis. While thereare no currently available approved vaccines, multivalent denguevaccines have shown some limited potential in humans (5, 12, 13, 28).However, vaccine development is difficult due to the presence of fourdistinct serotypes of the virus which each cause disease. Vaccinedevelopment also faces the challenge of ADE where unequal protectionagainst the different strains of the virus could actually increase therisk of more serious disease. Therefore there is a need for antiviraldrugs that target all of the serotypes of dengue. An antiviral drugadministered early during dengue infection that inhibits viralreplication would prevent the high viral load associated with DHF and bean attractive strategy in the treatment and prevention of disease. Anantiviral drug that inhibits viral replication could be administeredprior to travel to a dengue endemic region to prevent acquisition ofdisease, or for those that have previously been exposed to dengue, couldprevent infection by another serotype of virus and decrease the chanceof life-threatening DHF and DSS. Having an antiviral drug would also aidvaccine development by having a tool at hand to treat complications thatmay arise due to unequal immune protection against the differentserotypes. Although a successful vaccine could be a critical componentof an effective biodefense, the typical delay to onset of immunity,potential side-effects, cost, and logistics associated with large-scalecivilian vaccinations against a low-threat risk agent suggest that acomprehensive biodefense include a separate rapid-response element.Thus, there remains an urgent need to develop a safe and effectiveproduct to protect against flavivirus infection.

SUMMARY OF THE INVENTION

The present invention provides a pharmaceutical composition comprising apharmaceutically acceptable carrier and a compound having the followinggeneral formula I or a pharmaceutically acceptable salt thereof:

wherein R¹ and R² are independently hydrogen, alkyl, alkenyl, alkynyl,or unsubstituted or substituted cycloalkyl, arylalkyl, aryl, or R¹ andR² together may form a substituted or unsubstituted ring, which mayinclude one or more heteroatoms in the ring; and

Ar is substituted or unsubstituted aryl or heteroaryl;

said cycloalkyl, arylalkyl, and aryl group substituents being one ormore radical(s) independently selected from the group consisting of astraight- or branched chain alkyl, alkoxy, alkoxyalkyl, alkoxyalkoxy,halogen, polyfluoroalkyl, polyfluoroalkoxy, carboxy, cyano, nitro,amido, amidoalkyl, carboxamide, alkylthio, alkylsulfinyl, alkylsulfonyl,sulfonamide, and mercapto.

The present invention also provides a method for the treatment orprophylaxis of a viral infection or disease associated therewith,comprising administering in a therapeutically effective amount to amammal in need thereof, a compound of Formula I below or apharmaceutically acceptable salt thereof:

wherein R¹ and R² are independently hydrogen, alkyl, alkenyl, alkynyl,or unsubstituted or substituted cycloalkyl, arylalkyl, aryl, or R¹ andR² together may form a substituted or unsubstituted ring, which mayinclude one or more heteroatoms in the ring; and

Ar is substituted or unsubstituted aryl or heteroaryl;

said cycloalkyl, arylalkyl, and aryl group substituents being one ormore radical(s) independently selected from the group consisting of astraight- or branched chain alkyl, alkoxy, alkoxyalkyl, alkoxyalkoxy,halogen, polyfluoroalkyl, polyfluoroalkoxy, carboxy, cyano, nitro,amido, amidoalkyl, carboxamide, alkylthio, alkylsulfinyl, alkylsulfonyl,sulfonamide, and mercapto.

Other objects and advantages of the present invention will becomeapparent from the following description and appended claims.

DETAILED DESCRIPTION OF THE INVENTION

The compounds of the invention are of the following general Formula I:

wherein R¹ and R² are independently hydrogen, alkyl, alkenyl, alkynyl,or unsubstituted or substituted cycloalkyl, arylalkyl, aryl, or R¹ andR² together may form a substituted or unsubstituted ring, which mayinclude one or more heteroatoms in the ring; and

Ar is substituted or unsubstituted aryl or heteroaryl;

said cycloalkyl, arylalkyl, and aryl group substituents being one ormore radical(s) independently selected from the group consisting of astraight- or branched chain alkyl, alkoxy, alkoxyalkyl, alkoxyalkoxy,halogen, polyfluoroalkyl, polyfluoroalkoxy, carboxy, cyano, nitro,amido, amidoalkyl, carboxamide, alkylthio, alkylsulfinyl, alkylsulfonyl,sulfonamide, and mercapto.

Preferably each of R¹ and R² is an ethyl. Also preferably, Ar is amono-substituted phenyl such as amido-phenyl. Examples of amido-phenylinclude isobutyramidophenyl,p-[2-(4-oxo-4H-quinazolin-3-yl)-acetamido]-phenyl andp-[2-(4-oxo-4H-benzo[d][1,2,3]triazin-3-yl)-acetamido]-phenyl. Againpreferably, Ar is a di-substituted phenyl with one substituent as amidoand the other one as methoxy. Examples of this type of phenyl includem-phenyl-acetamido-p-methoxy-phenyl andm-(3-methyl-butyramido)-p-methoxy-phenyl.

Most preferably, the compound of Formula I isN-(4-Diethylsulfamoyl-phenyl)-2-(4-oxo-4H-quinazolin-3-yl)-acetamide.Exemplary compounds according to the invention are shown below in Table1.

The method of the present invention is for the treatment or prophylaxisof a viral infection or disease associated therewith, comprisingadministering in a therapeutically effective amount to a mammal in needthereof, a compound of Formula I described above.

Preferably, the mammal is a human and the viral infection is aflavivirus infection. More preferably, the flavivirus virus is selectedfrom the group consisting of Dengue virus, West Nile virus, yellow fevervirus, Japanese encephalitis virus, and tick-borne encephalitis virus.Most preferably, the flavivirus is a Dengue virus selected from thegroup consisting of DEN-1, DEN-2, DEN-3, and DEN-4.

Preferably, the viral infection is associated with a condition selectedfrom the group consisting of Dengue fever, Yellow fever, West Nile, St.Louis encephalitis, Hepatitis C, Murray Valley encephalitis, andJapanese encephalitis. Most preferably, the viral infection isassociated with Dengue fever wherein said Dengue fever is selected fromthe group consisting of classical dengue fever, dengue hemorrhagic feversyndrome, and dengue shock syndrome.

The method of the present invention may also comprise co-administrationof: a) other antivirals such as Ribavirin or cidofovir; b) vaccines;and/or c) interferons or pegylated interferons.

DEFINITIONS

In accordance with this detailed description, the followingabbreviations and definitions apply. It must be noted that as usedherein, the singular forms “a,” “an,” and “the” include plural referentsunless the context clearly dictates otherwise.

The publications discussed herein are provided solely for theirdisclosure. Nothing herein is to be construed as an admission regardingantedating the publications. Further, the dates of publication providedmay be different from the actual publication dates, which may need to beindependently confirmed.

Where a range of values is provided, it is understood that eachintervening value is encompassed. The upper and lower limits of thesesmaller ranges may independently be included in the smaller, subject toany specifically-excluded limit in the stated range. Where the statedrange includes one or both of the limits, ranges excluding either bothof those included limits are also included in the invention. Alsocontemplated are any values that fall within the cited ranges.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art. Any methods and materials similar or equivalent to thosedescribed herein can also be used in practice or testing. Allpublications mentioned herein are incorporated herein by reference todisclose and describe the methods and/or materials in connection withwhich the publications are cited.

By “patient” or “subject” is meant to include any mammal. A “mammal,”for purposes of treatment, refers to any animal classified as a mammal,including but not limited to, humans, experimental animals includingrats, mice, and guinea pigs, domestic and farm animals, and zoo, sports,or pet animals, such as dogs, horses, cats, cows, and the like.

The term “efficacy” as used herein in the context of a chronic dosageregime refers to the effectiveness of a particular treatment regime.Efficacy can be measured based on change of the course of the disease inresponse to an agent.

The term “success” as used herein in the context of a chronic treatmentregime refers to the effectiveness of a particular treatment regime.This includes a balance of efficacy, toxicity (e.g., side effects andpatient tolerance of a formulation or dosage unit), patient compliance,and the like. For a chronic administration regime to be considered“successful” it must balance different aspects of patient care andefficacy to produce a favorable patient outcome.

The terms “treating,” “treatment,” and the like are used herein to referto obtaining a desired pharmacological and physiological effect. Theeffect may be prophylactic in terms of preventing or partiallypreventing a disease, symptom, or condition thereof and/or may betherapeutic in terms of a partial or complete cure of a disease,condition, symptom, or adverse effect attributed to the disease. Theterm “treatment,” as used herein, covers any treatment of a disease in amammal, such as a human, and includes: (a) preventing the disease fromoccurring in a subject which may be predisposed to the disease but hasnot yet been diagnosed as having it, i.e., causing the clinical symptomsof the disease not to develop in a subject that may be predisposed tothe disease but does not yet experience or display symptoms of thedisease; (b) inhibiting the disease, i.e., arresting or reducing thedevelopment of the disease or its clinical symptoms; and (c) relievingthe disease, i.e., causing regression of the disease and/or its symptomsor conditions. Treating a patient's suffering from disease related topathological inflammation is contemplated. Preventing, inhibiting, orrelieving adverse effects attributed to pathological inflammation overlong periods of time and/or are such caused by the physiologicalresponses to inappropriate inflammation present in a biological systemover long periods of time are also contemplated.

“Alkenyl” refers to alkenyl group preferably having from 2 to 10 carbonatoms and more preferably 2 to 6 carbon atoms and having at least 1 andpreferably from 1-2 sites of alkenyl unsaturation.

“Alkoxy” refers to the group “alkyl-O—” which includes, by way ofexample, methoxy, ethoxy, n-propoxy, iso-propoxy, n-butoxy, tert-butoxy,sec-butoxy, n-pentoxy, n-hexoxy, 1,2-dimethylbutoxy, and the like.

“Alkyl” refers to linear or branched alkyl groups having from 1 to 10carbon atoms, alternatively 1 to 6 carbon atoms. This term isexemplified by groups such as methyl, t-butyl, n-heptyl, octyl and thelike. “Amino” refers to the group —NH₂.

“Aryl” or “Ar” refers to an unsaturated aromatic carbocyclic group offrom 6 to 14 carbon atoms having a single ring (e.g., phenyl) ormultiple condensed rings (e.g., naphthyl or anthryl) which condensedrings may or may not be aromatic (e.g., 2-benzoxazolinone,2H-1,4-benzoxazin-3(4H)-one, and the like) provided that the point ofattachment is through an aromatic ring atom. “Substituted aryl” refersto aryl groups which are substituted with from 1 to 3 substituentsselected from the group consisting of hydroxy, acyl, acylamino,thiocarbonylamino, acyloxy, alkyl, substituted alkyl, alkoxy,substituted alkoxy, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, amidino, alkylamidino, thioamidino, amino, aminoacyl,aminocarbonyloxy, aminocarbonylamino, aminothiocarbonylamino, aryl,substituted aryl, aryloxy, substituted aryloxy, cycloalkoxy, substitutedcycloalkoxy, heteroaryloxy, substituted heteroaryloxy, heterocyclyloxy,substituted heterocyclyloxy, carboxyl, carboxylalkyl,carboxyl-substituted alkyl, carboxyl-cycloalkyl, carboxyl-substitutedcycloalkyl, carboxylaryl, carboxyl-substituted aryl, carboxylheteroaryl,carboxyl-substituted heteroaryl, carboxylheterocyclic,carboxyl-substituted heterocyclic, carboxylamido, cyano, thiol,thioalkyl, substituted thioalkyl, thioaryl, substituted thioaryl,thioheteroaryl, substituted thioheteroaryl, thiocycloalkyl, substitutedthiocycloalkyl, thioheterocyclic, substituted thioheterocyclic,cycloalkyl, substituted cycloalkyl, guanidino, guanidinosulfone, halo,nitro, heteroaryl, substituted heteroaryl, heterocyclic, substitutedheterocyclic, cycloalkoxy, substituted cycloalkoxy, heteroaryloxy,substituted heteroaryloxy, heterocyclyloxy, substituted heterocyclyloxy,oxycarbonylamino, oxythiocarbonylamino, —S(O)₂-alkyl, —S(O)₂-substitutedalkyl, —S(O)₂-cycloalkyl, —S(O)₂-substituted cycloalkyl, —S(O)₂-alkenyl,—S(O)₂-substituted alkenyl, —S(O)₂-aryl, —S(O)₂-substituted aryl,—S(O)₂-heteroaryl, —S(O)₂-substituted heteroaryl, —S(O)₂-heterocyclic,—S(O)₂-substituted heterocyclic, —OS(O)₂-alkyl, —OS(O)₂-substitutedalkyl, —OS(O)₂-aryl, —OS(O)₂-substituted aryl, —OS(O)₂-heteroaryl,—OS(O)₂-substituted heteroaryl, —OS(O)₂-heterocyclic, —OS(O)₂—substituted heterocyclic, —OS(O)₂—NRR where R is hydrogen. or alkyl,—NRS(O)₂— alkyl, —NRS(O)₂-substituted alkyl, —NRS(O)₂-aryl,—NRS(O)₂-substituted aryl, —NRS(O)₂-heteroaryl, —NRS(O)₂-substitutedheteroaryl, —NRS(O)₂-heterocyclic, —NRS(O)₂-substituted heterocyclic,—NRS(O)₂—NR-alkyl, —NRS(O)₂—NR-substituted alkyl, —NRS(O)₂—NR-aryl,—NRS(O)₂—NR-substituted aryl, —NRS(O)₂—NR-heteroaryl,—NRS(O)₂—NR-substituted heteroaryl, —NRS(O)₂—NR-heterocyclic,—NRS(O)₂—NR-substituted heterocyclic where R is hydrogen or alkyl, mono-and di-alkylamino, mono- and di-(substituted alkyl)amino, mono- anddi-arylamino, mono- and di-substituted arylamino, mono- anddi-heteroarylamino, mono- and di-substituted heteroarylamino, mono- anddi-heterocyclic amino, mono- and di-substituted heterocyclic amino,unsymmetric di-substituted amines having different substituentsindependently selected from the group consisting of alkyl, substitutedalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl,heterocyclic and substituted heterocyclic and amino groups on thesubstituted aryl blocked by conventional blocking groups such as Boc,Cbz, formyl, and the like or substituted with —SO₂NRR where R ishydrogen or alkyl.

“Cycloalkyl” refers to cyclic alkyl groups of from 3 to 8 carbon atomshaving a single cyclic ring including, by way of example, cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl, cyclooctyl and the like. Excludedfrom this definition are multi-ring alkyl groups such as adamantanyl,etc.

“Halo” or “halogen” refers to fluoro, chloro, bromo and iodo.

“Heteroaryl” refers to an aromatic carbocyclic group of from 2 to 10carbon atoms and 1 to 4 heteroatoms selected from the group consistingof oxygen, nitrogen and sulfur within the ring or oxides thereof. Suchheteroaryl groups can have a single ring (e.g., pyridyl or furyl) ormultiple condensed rings (e.g., indolizinyl or benzothienyl) wherein oneor more of the condensed rings may or may not be aromatic provided thatthe point of attachment is through an aromatic ring atom. Additionally,the heteroatoms of the heteroaryl group may be oxidized, i.e., to formpyridine N-oxides or 1,1-dioxo-1,2,5-thiadiazoles and the like.Additionally, the carbon atoms of the ring may be substituted with anoxo (═O). The term “heteroaryl having two nitrogen atoms in theheteroaryl, ring” refers to a heteroaryl group having two, and only two,nitrogen atoms in the heteroaryl ring and optionally containing 1 or 2other heteroatoms in the heteroaryl ring, such as oxygen or sulfur.

“Substituted heteroaryl” refers to heteroaryl groups which aresubstituted with from 1 to 3 substituents selected from the groupconsisting of hydroxy, acyl, acylamino, thiocarbonylamino, acyloxy,alkyl, substituted alkyl, alkoxy, substituted alkoxy, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, amidino,alkylamidino, thioamidino, amino, aminoacyl, aminocarbonyloxy,aminocarbonylamino, aminothiocarbonylamino, aryl, substituted aryl,aryloxy, substituted aryloxy, cycloalkoxy, substituted cycloalkoxy,heteroaryloxy, substituted heteroaryloxy, heterocyclyloxy, substitutedheterocyclyloxy, carboxyl, carboxylalkyl, carboxyl-substituted alkyl,carboxyl-cycloalkyl, carboxyl-substituted cycloalkyl, carboxylaryl,carboxyl-substituted aryl, carboxylheteroaryl, carboxyl-substitutedheteroaryl, carboxylheterocyclic, carboxyl-substituted heterocyclic,carboxylamido, cyano, thiol, thioalkyl, substituted thioalkyl, thioaryl,substituted thioaryl, thioheteroaryl, substituted thioheteroaryl,thiocycloalkyl, substituted thiocycloalkyl, thioheterocyclic,substituted thioheterocyclic, cycloalkyl, substituted cycloalkyl,guanidino, guanidinosulfone, halo, nitro, heteroaryl, substitutedheteroaryl, heterocyclic, substituted heterocyclic, cycloalkoxy,substituted cycloalkoxy, heteroaryloxy, substituted heteroaryloxy,heterocyclyloxy, substituted heterocyclyloxy, oxycarbonylamino,oxythiocarbonylamino, —S(O)₂-alkyl, —S(O)₂-substituted alkyl,—S(O)₂-cycloalkyl, —S(O)₂-substituted cycloalkyl, —S(O)₂-alkenyl,—S(O)₂-substituted alkenyl, —S(O)₂-aryl, —S(O)₂-substituted aryl,—S(O)₂-heteroaryl, —S(O)₂-substituted heteroaryl, —S(O)₂-heterocyclic,—S(O)₂-substituted heterocyclic, —OS(O)₂-alkyl, —OS(O)₂-substitutedalkyl, —OS(O)₂-aryl, —OS(O)₂-substituted aryl, —OS(O)₂-heteroaryl,—OS(O)₂-substituted heteroaryl, —OS(O)₂-heterocyclic,—OS(O)₂-substituted heterocyclic, —OSO₂—NRR where R is hydrogen oralkyl, —NRS(O)₂-alkyl, —NRS(O)₂-substituted alkyl, —NRS(O)₂-aryl,—NRS(O)₂-substituted aryl, —NRS(O)₂-heteroaryl, —NRS(O)₂-substitutedheteroaryl, —NRS(O)₂-heterocyclic, —NRS(O)₂-substituted heterocyclic,—NRS(O)₂—NR-alkyl, —NRS(O)₂—NR-substituted alkyl, —NRS(O)₂—NR-aryl,—NRS(O)₂—NR-substituted aryl, —NRS(O)₂—NR-heteroaryl,—NRS(O)₂—NR-substituted heteroaryl, —NRS(O)₂—NR-heterocyclic,—NRS(O)₂—NR-substituted heterocyclic where R is hydrogen or alkyl, mono-and di-alkylamino, mono- and di-(substituted alkyl)amino, mono- anddi-arylamino, mono- and di-substituted arylamino, mono- anddi-heteroarylamino, mono- and di-substituted heteroarylamino, mono- anddi-heterocyclic amino, mono- and di-substituted heterocyclic amino,unsymmetric di-substituted amines having different substituentsindependently selected from the group consisting of alkyl, substitutedalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl,heterocyclic and substituted heterocyclic and amino groups on thesubstituted aryl blocked by conventional blocking groups such as Boc,Cbz, formyl, and the like or substituted with —SO₂NRR where R ishydrogen or alkyl.

“Sulfonyl” refers to the group —S(O)₂R where R is selected from thegroup consisting of hydrogen, alkyl, substituted alkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, aryl, substitutedaryl, cycloalkyl, substituted cycloalkyl, heteroaryl, substitutedheteroaryl, heterocyclic, substituted heterocyclic wherein alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl,heteroaryl, substituted heteroaryl, heterocyclic and substitutedheterocyclic are as defined herein.

“Pharmaceutically-acceptable carrier” means a carrier that is useful inpreparing a pharmaceutical composition or formulation that is generallysafe, non-toxic, and neither biologically nor otherwise undesirable, andincludes a carrier that is acceptable for veterinary use as well ashuman pharmaceutical use. A pharmaceutically-acceptable carrier orexcipient includes both one or more than one of such carriers.

“Pharmaceutically-acceptable cation” refers to the cation of apharmaceutically-acceptable salt. “Pharmaceutically-acceptable salt”refers to salts which retain the biological effectiveness and propertiesof compounds which are not biologically or otherwise undesirable.Pharmaceutically-acceptable salts refer to pharmaceutically-acceptablesalts of the compounds, which salts are derived from a variety oforganic and inorganic counter ions well known in the art and include, byway of example only, sodium, potassium, calcium, magnesium, ammonium,tetraalkylammonium, and the like; and when the molecule contains a basicfunctionality, salts of organic or inorganic acids, such ashydrochloride, hydrobromide, tartrate, mesylate, acetate, maleate,oxalate and the like.

Pharmaceutically-acceptable base addition salts can be prepared frominorganic and organic bases. Salts derived from inorganic bases, includeby way of example only, sodium, potassium, lithium, ammonium, calciumand magnesium salts. Salts derived from organic bases include, but arenot limited to, salts of primary, secondary and tertiary amines, such asalkyl amines, dialkyl amines, trialkyl amines, substituted alkyl amines,di(substituted alkyl) amines, tri(substituted alkyl) amines, alkenylamines, dialkenyl amines, trialkenyl amines, substituted alkenyl amines,di(substituted alkenyl) amines, tri(substituted alkenyl) amines,cycloalkyl amines, di(cycloalkyl) amines, tri(cycloalkyl) amines,substituted cycloalkyl amines, disubstituted cycloalkyl amine,trisubstituted cycloalkyl amines, cycloalkenyl amines, di(cycloalkenyl)amines, tri(cycloalkenyl) amines, substituted cycloalkenyl amines,disubstituted cycloalkenyl amine, trisubstituted cycloalkenyl amines,aryl amines, diaryl amines, triaryl amines, heteroaryl amines,diheteroaryl amines, triheteroaryl amines, heterocyclic amines,diheterocyclic amines, triheterocyclic amines, mixed di- and tri-amineswhere at least two of the substituents on the amine are different andare selected from the group consisting of alkyl, substituted alkyl,alkenyl, substituted alkenyl, cycloalkyl, substituted cycloalkyl,cycloalkenyl, substituted cycloalkenyl, aryl, heteroaryl, heterocyclic,and the like. Also included are amines where the two or threesubstituents, together with the amino nitrogen, form a heterocyclic orheteroaryl group.

Examples of suitable amines include, by way of example only,isopropylamine, trimethyl amine, diethyl amine, tri(iso-propyl) amine,tri(n-propyl) amine, ethanolamine, 2-dimethylaminoethanol, tromethamine,lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline,betaine, ethylenediamine, glucosamine, N-alkylglucamines, theobromine,purines, piperazine, piperidine, morpholine, N-ethylpiperidine, and thelike. It should also be understood that other carboxylic acidderivatives would be useful, for example, carboxylic acid amides,including carboxamides, lower alkyl carboxamides, dialkyl carboxamides,and the like.

Pharmaceutically-acceptable acid addition salts may be prepared frominorganic and organic acids. Salts derived from inorganic acids includehydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid,phosphoric acid, and the like. Salts derived from organic acids includeacetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid,malic acid, malonic acid, succinic acid, maleic acid, fumaric acid,tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid,methanesulfonic acid, ethanesulfonic acid, p-toluene-sulfonic acid,salicylic acid, and the like.

A compound may act as a pro-drug. Pro-drug means any compound whichreleases an active parent drug in vivo when such pro-drug isadministered to a mammalian subject. Pro-drugs are prepared by modifyingfunctional groups present in such a way that the modifications may becleaved in vivo to release the parent compound. Prodrugs includecompounds wherein a hydroxy, amino, or sulfhydryl group is bonded to anygroup that may be cleaved in vivo to regenerate the free hydroxyl,amino, or sulfhydryl group, respectively. Examples of prodrugs include,but are not limited to esters (e.g., acetate, formate, and benzoatederivatives), carbamates (e.g., N,N-dimethylamino-carbonyl) of hydroxyfunctional groups, and the like.

“Treating” or “treatment” of a disease includes:

(1) preventing the disease, i.e. causing the clinical symptoms of thedisease not to develop in a mammal that may be exposed to or predisposedto the disease but does not yet experience or display symptoms of thedisease,

(2) inhibiting the disease, i.e., arresting or reducing the developmentof the disease or its clinical symptoms, or

(3) relieving the disease, i.e., causing regression of the disease orits clinical symptoms.

A “therapeutically-effective amount” means the amount of a compoundthat, when administered to a mammal for treating a disease, issufficient to effect such treatment for the disease. The“therapeutically-effective amount” will vary depending on the compound,the disease, and its severity and the age, weight, etc., of the mammalto be treated.

Pharmaceutical Formulations of the Compounds

In general, compounds will be administered in atherapeutically-effective amount by any of the accepted modes ofadministration for these compounds. The compounds can be administered bya variety of routes, including, but not limited to, oral, parenteral(e.g., subcutaneous, subdural, intravenous, intramuscular, intrathecal,intraperitoneal, intracerebral, intraarterial, or intralesional routesof administration), topical, intranasal, localized (e.g., surgicalapplication or surgical suppository), rectal, and pulmonary (e.g.,aerosols, inhalation, or powder). Accordingly, these compounds areeffective as both injectable and oral compositions. The compounds can beadministered continuously by infusion or by bolus injection.

The actual amount of the compound, i.e., the active ingredient, willdepend on a number of factors, such as the severity of the disease,i.e., the condition or disease to be treated, age, and relative healthof the subject, the potency of the compound used, the route and form ofadministration, and other factors.

Toxicity and therapeutic efficacy of such compounds can be determined bystandard pharmaceutical procedures in cell cultures or experimentalanimals, e.g., for determining the LD₅₀ (the dose lethal to 50% of thepopulation) and the ED₅₀ (the dose therapeutically effective in 50% ofthe population). The dose ratio between toxic and therapeutic effects isthe therapeutic index and it can be expressed as the ratio LD₅₀/ED₅₀.

The data obtained from the cell culture assays and animal studies can beused in formulating a range of dosage for use in humans. The dosage ofsuch compounds lies within a range of circulating concentrations thatinclude the ED₅₀ with little or no toxicity. The dosage may vary withinthis range depending upon the dosage form employed and the route ofadministration utilized. For any compound used, thetherapeutically-effective dose can be estimated initially from cellculture assays. A dose may be formulated in animal models to achieve acirculating plasma concentration range which includes the IC₅₀ (i.e.,the concentration of the test compound which achieves a half-maximalinhibition of symptoms) as determined in cell culture. Such informationcan be used to more accurately determine useful doses in humans. Levelsin plasma may be measured, for example, by high performance liquidchromatography.

The amount of the pharmaceutical composition administered to the patientwill vary depending upon what is being administered, the purpose of theadministration, such as prophylaxis or therapy, the state of thepatient, the manner of administration, and the like. In therapeuticapplications, compositions are administered to a patient alreadysuffering from a disease in an amount sufficient to cure or at leastpartially arrest the symptoms of the disease and its complications. Anamount adequate to accomplish this is defined as“therapeutically-effective dose.” Amounts effective for this use willdepend on the disease condition being treated as well as by the judgmentof the attending clinician depending upon factors such as the severityof the inflammation, the age, weight, and general condition of thepatient, and the like.

The compositions administered to a patient are in the form of 24pharmaceutical compositions described supra. These compositions may besterilized by conventional sterilization techniques, or may be sterilefiltered. The resulting aqueous solutions may be packaged for use as is,or lyophilized, the lyophilized preparation being combined with asterile aqueous carrier prior to administration. It will be understoodthat use of certain of the foregoing excipients, carriers, orstabilizers will result in the formation of pharmaceutical salts.

The active compound is effective over a wide dosage range and isgenerally administered in a pharmaceutically- ortherapeutically-effective amount. The therapeutic dosage of thecompounds will vary according to, for example, the particular use forwhich the treatment is made, the manner of administration of thecompound, the health and condition of the patient, and the judgment ofthe prescribing physician. For example, for intravenous administration,the dose will typically be in the range of about 0.5 mg to about 100 mgper kilogram body weight. Effective doses can be extrapolated fromdose-response curves derived from in vitro or animal model test systems.Typically, the clinician will administer the compound until a dosage isreached that achieves the desired effect.

When employed as pharmaceuticals, the compounds are usually administeredin the form of pharmaceutical compositions. Pharmaceutical compositionscontain as the active ingredient one or more of the compounds above,associated with one or more pharmaceutically-acceptable carriers orexcipients. The excipient employed is typically one suitable foradministration to human subjects or other mammals. In making thecompositions, the active ingredient is usually mixed with an excipient,diluted by an excipient, or enclosed within a carrier which can be inthe form of a capsule, sachet, paper or other container. When theexcipient serves as a diluent, it can be a solid, semi-solid, or liquidmaterial, which acts as a vehicle, carrier, or medium for the activeingredient. Thus, the compositions can be in the form of tablets, pills,powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions,solutions, syrups, aerosols (as a solid or in a liquid medium),ointments containing, for example, up to 10% by weight of the activecompound, soft and hard gelatin capsules, suppositories, sterileinjectable solutions, and sterile packaged powders.

In preparing a formulation, it may be necessary to mill the activecompound to provide the appropriate particle size prior to combiningwith the other ingredients. If the active compound is substantiallyinsoluble, it ordinarily is milled to a particle size of less than 200mesh. If the active compound is substantially water soluble, theparticle size is normally adjusted by milling to provide a substantiallyuniform distribution in the formulation, e.g., about 40 mesh. Someexamples of suitable excipients include lactose, dextrose, sucrose,sorbitol, mannitol, starches, gum acacia, calcium phosphate, alginates,tragacanth, gelatin, calcium silicate, microcrystalline cellulose,polyvinylpyrrolidone, cellulose, sterile water, syrup, and methylcellulose. The formulations can additionally include: lubricating agentssuch as talc, magnesium stearate, and mineral oil; wetting agents;emulsifying and suspending agents; preserving agents such as methyl- andpropylhydroxy-benzoates; sweetening agents; and flavoring agents. Thecompositions of the invention can be formulated so as to provide quick,sustained, or delayed-release of the active ingredient afteradministration to the patient by employing procedures known in the art.

The quantity of active compound in the pharmaceutical composition andunit dosage form thereof may be varied or adjusted widely depending uponthe particular application, the manner or introduction, the potency ofthe particular compound, and the desired concentration. The term “unitdosage forms” refers to physically-discrete units suitable as unitarydosages for human subjects and other mammals, each unit containing apredetermined quantity of active material calculated to produce thedesired therapeutic effect, in association with a suitablepharmaceutical excipient.

The compound can be formulated for parenteral administration in asuitable inert carrier, such as a sterile physiological saline solution.The dose administered will be determined by route of administration.

Administration of therapeutic agents by intravenous formulation is wellknown in the pharmaceutical industry. An intravenous formulation shouldpossess certain qualities aside from being just a composition in whichthe therapeutic agent is soluble. For example, the formulation shouldpromote the overall stability of the active ingredient(s), also, themanufacture of the formulation should be cost-effective. All of thesefactors ultimately determine the overall success and usefulness of anintravenous formulation.

Other accessory additives that may be included in pharmaceuticalformulations and compounds as follow: solvents: ethanol, glycerol,propylene glycol; stabilizers: EDTA (ethylene diamine tetraacetic acid),citric acid; antimicrobial preservatives: benzyl alcohol, methylparaben, propyl paraben; buffering agents: citric acid/sodium citrate,potassium hydrogen tartrate, sodium hydrogen tartrate, aceticacid/sodium acetate, maleic acid/sodium maleate, sodium hydrogenphthalate, phosphoric acid/potassium dihydrogen phosphate, phosphoricacid/disodium hydrogen phosphate; and tonicity modifiers: sodiumchloride, mannitol, dextrose.

The presence of a buffer is necessary to maintain the aqueous pH in therange of from about 4 to about 8. The buffer system is generally amixture of a weak acid and a soluble salt thereof, e.g., sodiumcitrate/citric acid; or the monocation or dication salt of a dibasicacid, e.g., potassium hydrogen tartrate; sodium hydrogen tartrate,phosphoric acid/potassium dihydrogen phosphate, and phosphoricacid/disodium hydrogen phosphate.

The amount of buffer system used is dependent on (1) the desired pH; and(2) the amount of drug. Generally, the amount of buffer used is in a0.5:1 to 50:1 mole ratio of buffenalendronate (where the moles of bufferare taken as the combined moles of the buffer ingredients, e.g., sodiumcitrate and citric acid) of formulation to maintain a pH in the range of4 to 8 and generally, a 1:1 to 10:1 mole ratio of buffer (combined) todrug present is used.

A useful buffer is sodium citrate/citric acid in the range of 5 to 50 mgper ml. sodium citrate to 1 to 15 mg per ml. citric acid, sufficient tomaintain an aqueous pH of 4-6 of the composition.

The buffer agent may also be present to prevent the precipitation of thedrug through soluble metal complex formation with dissolved metal ions,e.g., Ca, Mg, Fe, Al, Ba, which may leach out of glass containers orrubber stoppers or be present in ordinary tap water. The agent may actas a competitive complexing agent with the drug and produce a solublemetal complex leading to the presence of undesirable particulates.

In addition, the presence of an agent, e.g., sodium chloride in anamount of about of 1-8 mg/ml, to adjust the tonicity to the same valueof human blood may be required to avoid the swelling or shrinkage oferythrocytes upon administration of the intravenous formulation leadingto undesirable side effects such as nausea or diarrhea and possibly toassociated blood disorders. In general, the tonicity of the formulationmatches that of human blood which is in the range of 282 to 288 mOsm/kg,and in general is 285 mOsm/kg, which is equivalent to the osmoticpressure corresponding to a 0.9% solution of sodium chloride.

An intravenous formulation can be administered by direct intravenousinjection, i.v. bolus, or can be administered by infusion by addition toan appropriate infusion solution such as 0.9% sodium chloride injectionor other compatible infusion solution.

The compositions are preferably formulated in a unit dosage form, eachdosage containing from about 5 to about 100 mg, more usually about 10 toabout 30 mg, of the active ingredient. The term “unit dosage forms”refers to physically discrete units suitable as unitary dosages forhuman subjects and other mammals, each unit containing a predeterminedquantity of active material calculated to produce the desiredtherapeutic effect, in association with a suitable pharmaceuticalexcipient.

The active compound is effective over a wide dosage range and isgenerally administered in a pharmaceutically effective amount. It willbe understood, however, that the amount of the compound actuallyadministered will be determined by a physician, in the light of therelevant circumstances, including the condition to be treated, thechosen route of administration, the actual compound administered, theage, weight, and response of the individual patient, the severity of thepatient's symptoms, and the like.

For preparing solid compositions such as tablets, the principal activeingredient is mixed with a pharmaceutical excipient to form a solidpreformulation composition containing a homogeneous mixture of acompound of the present invention. When referring to thesepreformulation compositions as homogeneous, it is meant that the activeingredient is dispersed evenly throughout the composition so that thecomposition may be readily subdivided into equally effective unit dosageforms such as tablets, pills and capsules. This solid preformulation isthen subdivided into unit dosage forms of the type described abovecontaining from, for example, 0.1 to about 2000 mg of the activeingredient.

The tablets or pills may be coated or otherwise compounded to provide adosage form affording the advantage of prolonged action. For example,the tablet or pill can comprise an inner dosage and an outer dosagecomponent, the latter being in the form of an envelope over the former.The two components can be separated by an enteric layer which serves toresist disintegration in the stomach and permit the inner component topass intact into the duodenum or to be delayed in release. A variety ofmaterials can be used for such enteric layers or coatings, suchmaterials including a number of polymeric acids and mixtures ofpolymeric acids with such materials as shellac, cetyl alcohol, andcellulose acetate.

The liquid forms in which the novel compositions may be incorporated foradministration orally or by injection include aqueous solutions suitablyflavored syrups, aqueous or oil suspensions, and flavored emulsions withedible oils such as cottonseed oil, sesame oil, coconut oil, or peanutoil, as well as elixirs and similar pharmaceutical vehicles.

Compositions for inhalation or insufflation include solutions andsuspensions in pharmaceutically-acceptable, aqueous or organic solvents,or mixtures thereof, and powders. The liquid or solid compositions maycontain suitable pharmaceutically-acceptable excipients as describedsupra. Compositions in pharmaceutically-acceptable solvents may benebulized by use of inert gases. Nebulized solutions may be breatheddirectly from the nebulizing device or the nebulizing device may beattached to a face masks tent, or intermittent positive pressurebreathing machine. Solution, suspension, or powder compositions may beadministered from devices which deliver the formulation in anappropriate manner.

The compounds can be administered in a sustained release form. Suitableexamples of sustained-release preparations include semipermeablematrices of solid hydrophobic polymers containing the compounds, whichmatrices are in the form of shaped articles, e.g., films, ormicrocapsules. Examples of sustained-release matrices includepolyesters, hydrogels (e.g., poly(2-hydroxyethyl-methacrylate) asdescribed by Langer et al., J. Biomed. Mater. Res. 15: 167-277 (1981)and Langer, Chem. Tech. 12: 98-105 (1982) or poly(vinyl alcohol)),polylactides (U.S. Pat. No. 3,773,919), copolymers of L-glutamic acidand gamma ethyl-L-glutamate (Sidman et al., Biopolymers 22: 547-556,1983), non-degradable ethylene-vinyl acetate (Langer et al., supra),degradable lactic acid-glycolic acid copolymers such as the LUPRONDEPOT™ (i.e., injectable microspheres composed of lactic acid-glycolicacid copolymer and leuprolide acetate), and poly-D-(−)-3-hydroxybutyricacid (EP 133,988).

The compounds can be administered in a sustained-release form, forexample a depot injection, implant preparation, or osmotic pump, whichcan be formulated in such a manner as to permit a sustained-release ofthe active ingredient. Implants for sustained-release formulations arewell-known in the art. Implants may be formulated as, including but notlimited to, microspheres, slabs, with biodegradable or non-biodegradablepolymers. For example, polymers of lactic acid and/or glycolic acid forman erodible polymer that is well-tolerated by the host.

Transdermal delivery devices (“patches”) may also be employed. Suchtransdermal patches may be used to provide continuous or discontinuousinfusion of the compounds in controlled amounts. The construction anduse of transdermal patches for the delivery of pharmaceutical agents iswell known in the art. See, e.g., U.S. Pat. No. 5,023,252, issued Jun.11, 1991, herein incorporated by reference. Such patches may beconstructed for continuous, pulsatile, or on-demand delivery ofpharmaceutical agents.

Direct or indirect placement techniques may be used when it is desirableor necessary to introduce the pharmaceutical composition to the brain.Direct techniques usually involve placement of a drug delivery catheterinto the host's ventricular system to bypass the blood-brain barrier.One such implantable delivery system used for the transport ofbiological factors to specific anatomical regions of the body isdescribed in U.S. Pat. No. 5,011,472, which is herein incorporated byreference. Indirect techniques usually involve formulating thecompositions to provide for drug latentiation by the conversion ofhydrophilic drugs into lipid-soluble drugs. Latentiation is generallyachieved through blocking of the hydroxy, carbonyl, sulfate, and primaryamine groups present on the drug to render the drug more lipid-solubleand amenable to transportation across the blood-brain barrier.Alternatively, the delivery of hydrophilic drugs may be enhanced byintra-arterial infusion of hypertonic solutions which can transientlyopen the blood-brain barrier.

In order to enhance serum half-life, the compounds may be encapsulated,introduced into the lumen of liposomes, prepared as a colloid, or otherconventional techniques may be employed which provide an extended serumhalf-life of the compounds. A variety of methods are available forpreparing liposomes, as described in, e.g., Szoka et al., U.S. Pat. Nos.4,235,871, 4,501,728 and 4,837,028 each of which is incorporated hereinby reference.

Pharmaceutical compositions are suitable for use in a variety of drugdelivery systems. Suitable formulations for use in the present inventionare found in Remington's Pharmaceutical Sciences, Mace PublishingCompany, Philadelphia, Pa., 17th ed. (1985).

The provided compounds and pharmaceutical compositions show biologicalactivity in treating and preventing viral infections and associateddiseases, and, accordingly, have utility in treating viral infectionsand associated diseases, such as Hemorrhagic fever viruses, in mammalsincluding humans.

Hemorrhagic fever viruses (HFVs) are RNA viruses that cause a variety ofdisease syndromes with similar clinical characteristics. HFVs that areof concern as potential biological weapons include but are not limitedto: Arenaviridae (Junin, Machupo, Guanarito, Sabia, and Lassa),Filoviridae (Ebola and Marburg viruses), Flaviviridae (yellow fever,Omsk hemorrhagic fever and Kyasanur Forest disease viruses), andBunyaviridae (Rift Valley fever and Crimean-Congo hemorrhagic fever).The naturally-occurring arenaviruses and potential engineeredarenaviruses are included in the Category A Pathogen list according tothe Centers for Disease Control and Prevention as being among thoseagents that have greatest potential for mass casualties.

Risk factors include: travel to Africa or Asia, handling of animalcarcasses, contact with infected animals or people, and/or arthropodbites. Arenaviruses are highly infectious after direct contact withinfected blood and/or bodily secretions. Humans usually become infectedthrough contact with infected rodents, the bite of an infectedarthropod, direct contact with animal carcasses, inhalation ofinfectious rodent excreta and/or injection of food contaminated withrodent excreta. The Tacaribe virus has been associated with bats.Airborne transmission of hemorrhagic fever is another mode.Person-to-person contact may also occur in some cases.

All of the hemorrhagic fevers exhibit similar clinical symptoms.However, in general the clinical manifestations are non-specific andvariable. The incubation period is approximately 7-14 days. The onset isgradual with fever and malaise, tachypnea, relative bradycardia,hypotension, circulatory shock, conjunctival infection, pharyngitis,lymphadenopathy, encephalitis, myalgia, back pain, headache anddizziness, as well as hyperesthesia of the skin. Some infected patientsmay not develop hemorrhagic manifestations.

Methods of diagnosis at specialized laboratories include antigendetection by antigen-capture enzyme-linked immunosorbent assay (ELISA),IgM antibody detection by antibody-capture enzyme-linked immunosorbentassay, reverse transcriptase polymerase chain reaction (RT-PCR), andviral isolation. Antigen detection (by enzyme-linked immunosorbentassay) and reverse transcriptase polymerase chain reaction are the mostuseful diagnostic techniques in the acute clinical setting. Viralisolation is of limited value because it requires a biosafety level 4(BSL-4) laboratory.

Example 1 Determining Anti Dengue-2 Activity of Compounds of theInvention

A sensitive and reproducible high-throughput screening (HTS) assay hasbeen established to measure dengue virus-induced cytopathic effect(CPE). To determine the amount of dengue virus stock required to producecomplete CPE in 5 days, Vero cell monolayers were seeded on 96-wellplates and infected with 10-fold serial dilutions of the dengue virusstock representing a multiplicity of infection (MOI) of approximately0.001 PFU/cell to 0.1 PFU/cell. At 5 days post-infection, the cultureswere fixed with 5%, glutaraldehyde and stained with 0.1% crystal violet.Virus-induced CPE was quantified spectrophometrically at OD₅₇₀. Fromthis analysis, an MOI of 0.1 PFU/cell of dengue virus stock was chosenfor use in the HTS assay. To establish the signal-to-noise ratio (S/N)of the 96-well assay and evaluate the well-to-well and assay-to-assayvariability, five independent experiments were performed. Vero cellmonolayers were infected with 0.1 PFU/cell of dengue virus stock. Eachplate contained the following controls: quadruplicate virus-infectedwells, quadruplicate uninfected cell wells and a dose response curve induplicate for ribavirin at 500, 250, 125 and 62 μM, as referencestandards. At day 5 post-infection, the plates were processed asdescribed above.

Compounds were dissolved in DMSO and diluted in medium such that thefinal concentration in each well was 5 μM compound and 0.5% DMSO. Thecompounds were added robotically to the culture medium using thePerkinElmer MultiPROBE® II HT PLUS robotic system. Following compoundaddition, cultures were infected with dengue virus (DEN-2 strain NewGuinea C). After 5 days incubation, plates were processed and CPEquantified on a PerkinElmer EnVision II plate reader system.

The results of these experiments indicated that the 96-well assay formatis robust and reproducible. The S/N ratio (ratio of signal of cellcontrol wells (signal) to virus control wells (noise)) was 5.0±1.2. Thewell-to-well variability was determined for each individual plate andfound to have a coefficient of variance of less than 10% for bothpositive control and negative control wells, and overall assay-to-assayvariability was less than 15%. Using this assay, the EC₅₀ values forribavirin were determined to be 125±25 μM, respectively. Theeffectiveness of ribavirin against dengue varies with the cell typeused, but the values obtained were within the range of published valuesfor this compound (2, 14, 32). Taken together, these results show that asensitive and reproducible HTS assay has been successfully developed toevaluate our compound library for inhibitors of dengue virusreplication.

This assay was the basis of a high-throughput screen for dengue virusinhibitors, against which a library of 210,000 compounds was tested.Compounds that inhibited dengue virus induced CPE by at least 50% werefurther investigated for chemical tractability, potency, andselectivity.

Initially, the chemical structures of the hit compounds were examinedfor chemical tractability. A chemically tractable compound is defined asone that is synthetically accessible using reasonable chemicalmethodology, and which possesses chemically stable functionalities andpotential drug-like qualities. Hits that passed this medicinal chemistryfilter were evaluated for their potency. Compound potency was determinedby evaluating inhibitory activity across a broad range ofconcentrations. Nonlinear regression was used to generate best-fitinhibition curves and to calculate the 50% effective concentration(EC50).

Compounds that were active in the primary screen were also tested foractivity in viral yield assays. Table 1 below shows some of thecompounds that were tested for activity against Dengue-2 (Strain NewGuinea C) in a viral yield assay at a range of concentrations. Verocells in 12-well plates were infected with dengue-2 virus at amultiplicity of infection (MOI) of 0.1, treated with compound (or DMSOas a control), incubated at 37° C., harvested 48 hours post infectionand titered on Vero cells as described above. The EC50 was calculatedthrough ExcelFit. Those compounds with activity below 1 μM are indicatedwith “A”, those with activity between 1 and 10 μM are indicated with“B”, those with activity between 10 and 25 μM with “C”, and those withactivity above 25 μM are indicated with “D”.

TABLE 1 List of compounds of the present invention and their anti-dengue2 viral activity. Activity A: EC₅₀ < 1 uM; Molec- B: 1< EC₅₀ < 10 uM;Com- ular C: 10 < EC₅₀ < 25 uM; pound Chemical Structure Weight ChemicalName D: EC₅₀ > 25 uM 1

414.5 N-(4- Diethylsulfamoyl- phenyl)-2-(4-oxo- 4H-quinazolin-3-yl)-acetamide A 2

328.4 N-(5- Diethylsulfamoyl- 2-methoxy- phenyl)- isobutyramide A 3

415.5 N-(4- Diethylsulfamoyl- phenyl)-2-(4-oxo-4H-benzo[d][1,2,3]triazin- 3-yl)-acetamide B 4

448.5 N-(4- Diethylsulfamoyl- phenyl)-2-(1,3- dimethyl-2,6-dioxo-1,2,3,6- tetrahydro-purin- 7-yl)-acetamide B 5

420.5 N-(4- Diethylsulfamoyl- phenyl)-2-(4-oxo- 4H-thieno[2,3-d]pyrimidin-3- yl)-acetamide B 6

448.9 2-(6-Chloro-4- oxo-4H- quinazolin-3- yl)-N-[4- (isopropyl-methyl-sulfamoyl)-phenyl]- acetamide B 7

432.5 2-(7-Fluoro-4-oxo- 4H-quinazolin- 3-yl)-N-[4- (isopropyl-methyl-sulfamoyl)-phenyl]- acetamide B 8

430.5 N-(4- Diethylsulfamoyl- phenyl)-2-(1,4- dioxo-3,4- dihydro-1H-phthalazin-2-yl)- acetamide B 9

465.5 N-(4- Diethylsulfamoyl- phenyl)-2-(1,3- dioxo-1H,3H- benzo[de]isoquinolin-2- yl)-acetamide B 10

448.6 N-(4- Diethylsulfamoyl- phenyl)-2- (5,6-dimethyl-4- oxo-4H-thieno[2,3- d]pyrimidin-3-yl)- acetamide B 11

448.6 N-(4- Diethylsulfamoyl- phenyl)-2-(6- ethyl-4-oxo-4H- thieno[2,3-d]pyrimidin- 3-yl)-acetamide B 12

473.5 N-(5- Diethylsulfamoyl- 2-methyl- phenyl)-2-(7- nitro-4-oxo-4H-quinazolin-3-yl)- acetamide B 13

478.5 N-(5- Diethylsulfamoyl- 2-methoxy-phenyl)- 2-(1,3-dimethyl-2,6-dioxo-1,2,3,6- tetrahydro-purin- 7-yl)-acetamide B 14

328.4 N-(5- Diethylsulfamoyl- 2-methoxy- phenyl)- butyramide B 15

342.5 N-(5- Diethylsulfamoyl- 2-methoxy- phenyl)-3-methyl- butyramide B16

376.5 N-(5- Diethylsulfamoyl- 2-methoxy- phenyl)-2-phenyl- acetamide B17

440.5 1-[2-(4-Oxo-4H- quinazolin-3-yl)- acetyl]-2,3-dihydro-1H-indole-5- sulfonic acid diethylamide B 18

454.5 N-[4-(2,6- Dimethyl- piperidine-1- sulfonyl)-phenyl]- 2-(4-oxo-4H-quinazolin-3-yl)- acetamide B 19

454.5 N-[4-(3,5- Dimethyl- piperidine-1- sulfonyl)-phenyl]- 2-(4-oxo-4H-quinazolin-3- yl)-acetamide B 20

445.5 N-(5- Diethylsulfamoyl- 2-methoxy- phenyl)-2-(4-oxo-4H-benzo[d][1,2,3]triazin- 3-yl)-acetamide B 21

342.5 Pentanoic acid (5- diethylsulfamoyl- 2-methoxy- phenyl)-amide C 22

420.5 N-(4- Diethylsulfamoyl- phenyl)-2-(4-oxo- 4H-thieno[3,2-d]pyrimidin-3- yl)-acetamide C 23

448.9 2-(7-Chloro-4-oxo- 4H-quinazolin- 3-yl)-N-(4- diethylsulfamoyl-phenyl)-acetamide C 24

454.5 N-(4- Diethylsulfamoyl- phenyl)-2-(4-oxo- 4H-benzo [4,5]furo[3,2-d]pyrimidin-3- yl)-acetamide C 25

487.6 N-(5- Diethylsulfamoyl-2- isopropoxy-phenyl)- 3-(4-oxo-4H-benzo[d][1,2,3]triazin- 3-yl)-propionamide C 26

428.5 N-(4- Diethylsulfamoyl- phenyl)-2-(8- methyl-4-oxo- 4H-quinazolin-3-yl)-acetamide C 27

459.5 N-(5- Diethylsulfamoyl-2- methoxy-phenyl)- 3-(4-oxo-4H-benzo[d][1,2,3]triazin- 3-yl)- propionamide C 28

460.6 N-[4-(2,6- Dimethyl- piperidine-1- sulfonyl)-phenyl]- 2-(4-oxo-4H-thieno[2,3- d]pyrimidin-3- yl)-acetamide C 29

471.6 N-(4- Diethylsulfamoyl- phenyl)-2-[ethyl-(4- oxo-3,4-dihydro-quinazolin-2- ylmethyl)-amino]- acetamide C 30

440.4 2-(4-Oxo-4H- quinazolin-3-yl)- N-[4-(2,2,2- trifluoro-ethylsulfamoyl)- phenyl]-acetamide C 31

412.5 2-(4-Oxo-4H- quinazolin-3-yl)- N-[4-(pyrrolidine-1-sulfonyl)-phenyl]- acetamide D 32

441.5 2-(4-Oxo-4H- quinazolin-3-yl)- N-[4-(thiazol-2- ylsulfamoyl)-phenyl]-acetamide D 33

464.5 N-[4-(4,6-Dimethyl- pyrimidin-2- ylsulfamoyl)- phenyl]-2-(4-oxo-4H-quinazolin- 3-yl)-acetamide D 34

428.5 N-[4-(Morpholine- 4-sulfonyl)- phenyl]-2- (4-oxo-4H-quinazolin-3-yl)- acetamide D 35

428.5 N-[4-(Butyl-methyl- sulfamoyl)-phenyl]- 2-(4-oxo-4H-quinazolin-3-yl)- acetamide D 36

462.5 N-[4-(Benzyl- methyl-sulfamoyl)- phenyl]-2-(4-oxo-4H-quinazolin-3- yl)-acetamide D 37

440.5 N-[4-(2-Methyl- piperidine-1- sulfonyl)-phenyl]- 2-(4-oxo-4H-quinazolin-3-yl)- acetamide D 38

358.4 2-(4-Oxo-4H- quinazolin-3-yl)- N-(4-sulfamoyl- phenyl)-acetamide D39

436.5 2-(4-Oxo-4H- quinazolin-3-yl)- N-[4-(pyrimidin- 2-ylsulfamoyl)-phenyl]-acetamide D 40

485.6 N-[4-(3- Morpholin-4-yl- propylsulfamoyl)- phenyl]-2-(4-oxo-4H-quinazolin- 3-yl)-acetamide D 41

438.5 N-{4-[(Furan- 2-ylmethyl)- sulfamoyl]- phenyl}-2-(4- oxo-4H-quinazolin-3- yl)-acetamide D 42

414.5 N-[4-(Isopropyl- methyl-sulfamoyl)- phenyl]-2-(4-oxo-4H-quinazolin- 3-yl)-acetamide D 43

480.6 N-[4-(1- Bicyclo[2.2.1]hept- 2-yl- ethylsulfamoyl)-phenyl]-2-(4-oxo- 4H-quinazolin- 3-yl)-acetamide D 44

400.5 N-(4- Isopropyl- sulfamoyl- phenyl)-2-(4-oxo- 4H-quinazolin-3-yl)-acetamide D 45

441.5 N-[4-(4-Methyl- piperazine-1- sulfonyl)-phenyl]- 2-(4-oxo-4H-quinazolin-3-yl)- acetamide D 46

372.4 N-(4- Methylsulfamoyl- phenyl)-2-(4-oxo- 4H-quinazolin-3-yl)-acetamide D 47

440.5 N-[4-(4-Methyl- piperidine-1- sulfonyl)-phenyl]- 2-(4-oxo-4H-quinazolin-3- yl)-acetamide D 48

476.5 N-[4-(1,1-Dioxo- tetrahydro- 1lambda*6*- thiophen-3- ylsulfamoyl)-phenyl]- 2-(4-oxo-4H- quinazolin-3-yl)- acetamide D 49

454.5 N-[4-(Cyclohexyl- methyl-sulfamoyl)- phenyl]-2-(4-oxo-4H-quinazolin- 3-yl)-acetamide D 50

386.4 N-(4- Dimethylsulfamoyl- phenyl)-2-(4-oxo- 4H-quinazolin-3-yl)-acetamide D 51

440.5 N-[4-(Azepane- 1-sulfonyl)- phenyl]-2-(4-oxo- 4H-quinazolin-3-yl)-acetamide D 52

426.5 2-(4-Oxo-4H- quinazolin-3-yl)- N-[4-(piperidine-1-sulfonyl)-phenyl]- acetamide D 53

440.5 N-[4-(3-Methyl- piperidine-1- sulfonyl)-phenyl]- 2-(4-oxo-4H-quinazolin-3- yl)-acetamide D 54

430.4 {4-[2-(4-Oxo- 4H-quinazolin- 3-yl)-acetylamino]- benzene-sulfonylamino}- acetic acid methyl ester D 55

414.5 N-(3-tert- Butylsulfamoyl- phenyl)-2-(4-oxo- 4H-quinazolin-3-yl)-acetamide D 56

496.6 N-(4- Diethylsulfamoyl- phenyl)-2-(4-oxo- 6-phenyl-4H- thieno[2,3-d]pyrimidin- 3-yl)-acetamide D 57

432.5 N-(4- Diethylsulfamoyl- phenyl)-2-(7-fluoro- 4-oxo-4H-quinazolin-3-yl)- acetamide D 58

474.5 N-(4- Diethylsulfamoyl- phenyl)-2-(6,7- dimethoxy-4-oxo-4H-quinazolin- 3-yl)-acetamicle D 59

496.6 N-(4- Diethylsulfamoyl- phenyl)-2-(4-oxo- 5-phenyl-4H- thieno[2,3-d]pyrimidin- 3-yl)-acetamide D 60

488.5 N-(4- Diethylsulfamoyl- phenyl)-2-(7- methylamino-6-nitro-4-oxo-4H- quinazolin-3-yl)- acetamide D 61

526.6 N-(4- Diethylsulfamoyl- phenyl)-2-[6-(4- methoxy-phenyl)-4-oxo-4H- thieno[3,2- d]pyrimidin- 3-yl]-acetamide D 62

400.5 N-(4- Dimethylsulfamoyl- phenyl)-2-(8- methyl-4-oxo-4H-quinazolin-3- yl)-acetamide D 63

417.5 N-[(4- Diethylsulfamoyl- phenylcarbamoyl)- methyl]-3,5- dimethyl-benzamide D 64

474.6 N-(4- Diethylsulfamoyl- phenyl)-2-(3- ethyl-4-oxo-3,4-dihydro-quinazolin- 2-ylsulfanyl)- acetamide D 65

414.5 N-(3- Diethylsulfamoyl- phenyl)-2-(4- oxo-4H-quinazolin-3-yl)-acetamide D 66

472.5 4-Diethylsulfamoyl- benzoic acid 3- carbamoylmethyl-4-oxo-3,4-dihydro- quinazolin-2- ylmethyl ester D 67

400.5 4-Diethylsulfamoyl- N-(4-oxo-4H- quinazolin-3- yl)-benzamide D 68

457.6 N-[1-(4- Diethylsulfamoyl- phenyl)-ethyl]- 3-(4-oxo-4H-benzo[d][1,2,3]triazin- 3-yl)-propionamide D 69

428.5 N-(4- Diethylsulfamoyl- benzyl)-2-(4-oxo- 4H-quinazolin-3-yl)-acetamide D 70

496.6 N-[(4- Diethylsulfamoyl- phenylcarbamoyl)- methyl]-3-dimethylsulfamoyl- benzamide D 71

460.6 N-(4- Diethylsulfamoyl- phenyl)-2-(3- methyl-4-oxo-3,4-dihydro-quinazolin- 2-ylsulfanyl)- acetamide D 72

403.5 N-[(4- Diethylsulfamoyl- phenylcarbamoyl)- methyl]-4-methyl-benzamide D 73

510.6 N-[(4- Diethylsulfamoyl- phenylcarbamoyl)- methyl]-4-isopropylsulfamoyl- benzamide D 74

439.5 Naphthalene-2- carboxylic acid[(4- diethylsulfamoyl-phenylcarbamoyl)- methyl]-amide D 75

506.0 2-[(7-Chloro- 4-oxo-3,4- dihydro-quinazolin- 2-ylmethyl)-ethyl-amino]-N-(4- diethylsulfamoyl- phenyl)-acetamide D 76

460.6 4-{[(4- Diethylsulfamoyl- phenylcarbamoyl)- methyl]-amino}-N,N-diethyl- benzamide D 77

429.5 N-(4- Diethylsulfamoyl- phenyl)-3- (4-oxo-4H-benzo[d][1,2,3]triazin- 3-yl)- propionamide D 78

458.5 N-(4- Diethylsulfamoyl- phenyl)-2-(3- ethyl-2,4-dioxo-3,4-dihydro-2H- quinazolin-1-yl)- acetamide D 79

449.5 N-[(4- Diethylsulfamoyl- phenylcarbamoyl)- methyl]-3,4- dimethoxy-benzamide D 80

465.6 Biphenyl-4- carboxylic acid [(4- diethylsulfamoyl-phenylcarbamoyl)- methyl]-amide D 81

473.5 (4-Oxo-4H- benzo[d][1,2,3]triazin- 3-yl)-acetic acid (4-diethylsulfamoyl- phenylcarbamoyl)- methyl ester D 82

429.5 N-[4- (Morpholine-4- sulfonyl)-phenyl]- 2-(4-oxo-4H-benzo[d][1,2,3]triazin- 3-yl)-acetamide D 83

481.6 N-[4-(1- Bicyclo[2.2.1]hept- 2-yl-ethylsulfamoyl)-phenyl]-2-(4-oxo-4H- benzo[d][1,2,3]triazin- 3-yl)-acetamide D 84

403.4 N-[4- (Methoxy-methyl- sulfamoyl)-phenyl]- 2-(4-oxo-4H-benzo[d][1,2,3]triazin- 3-yl)-acetamide D 85

387.4 N-(4- Dimethylsulfamoyl- phenyl)-2-(4-oxo-4H-benzo[d][1,2,3]triazin- 3-yl)-acetamide D 86

415.5 N-[4- (Isopropyl-methyl- sulfamoyl)-phenyl]- 2-(4-oxo-4H-benzo[d][1,2,3]triazin- 3-yl)-acetamide D 87

427.5 2-(4-Oxo-4H- benzo[d][1,2,3]triazin- 3-yl)-N-[4- (piperidine-1-sulfonyl)-phenyl]- acetamide D 88

457.6 N-(5- Diethylsulfamoyl- 2-dimethylamino- phenyl)-2-(4-oxo-4H-quinazolin-3- yl)-acetamide D 89

428.5 N-(5- Diethylsulfamoyl- 2-methyl-phenyl)- 2-(4-oxo-4H-quinazolin-3- yl)-acetamide D 90

442.5 N-(5- Diethylsulfamoyl- 2,3-dimethyl- phenyl)-2-(4-oxo-4H-quinazolin-3- yl)-acetamide D 91

444.5 N-(5- Diethylsulfamoyl- 2-methoxy- phenyl)-2-(4-oxo-4H-quinazolin-3- yl)-acetamide D 92

458.5 N-(5- Diethylsulfamoyl- 2-ethoxy-phenyl)- 2-(4-oxo-4H-quinazolin-3- yl)-acetamide D 93

448.9 N-(2-Chloro-5- diethylsulfamoyl- phenyl)-2-(4-oxo- 4H-quinazolin-3-yl)-acetamide D 94

431.5 N-(5- Diethylsulfamoyl- 2-hydroxy-phenyl)- 2-(4-oxo-4H-benzo[d][1,2,3]triazin- 3-yl)-acetamide D 95

429.5 N-(5- Diethylsulfamoyl- 2-methyl-phenyl)- 2-(4-oxo-4H-benzo[d][1,2,3]triazin- 3-yl)-acetamide D 96

426.5 1-(3-Methyl-4- oxo-3,4-dihydro- quinazolin-2- ylmethyl)-2,3-dihydro-1H- indole-5-sulfonic acid diethylamide D 97

468.6 1-[2-(8-Methyl-4- oxo-4H-quinazolin- 3-yl)-acetyl]-1,2,3,4-tetrahydro- quinoline-6- sulfonic acid diethylamide D 98

476.6 N-[4-(Benzyl- methyl-sulfamoyl)- phenyl]-3-(4-oxo-4H-quinazolin-3- yl)-propionamide D 99

405.5 N-(3-Benzyl-4- oxo-3,4-dihydro- quinazolin-6-yl)- 4-methyl-benzenesulfonamide D 100

372.4 2-(4-Methyl-1-oxo- 1H-phthalazin-2-yl)- N-(4-sulfamoyl-phenyl)-acetamide D 101

430.5 2-(3-Ethyl-2,4- dioxo-3,4-dihydro- 2H-quinazolin-1- yl)-N-[2-(4-sulfamoyl-phenyl)- ethyl]-acetamide D 102

416.5 2-(3-Methyl-2,4- dioxo-3,4-dihydro- 2H-quinazolin-1- yl)-N-[2-(4-sulfamoyl-phenyl)- ethyl]-acetamide D 103

400.5 N-{2-[4-(3- Methyl-4-oxo-3,4- dihydro-quinazolin- 6-ylsulfamoyl)-phenyl]-ethyl}- acetamide D 104

476.6 N-{2-[4-(3- Benzyl-4-oxo-3,4- dihydro-quinazolin- 6-ylsulfamoyl)-phenyl]-ethyl}- acetamide D 105

428.5 N-(3- Diethylsulfamoyl- 4-methyl-phenyl)- 2-(4-oxo-4H-quinazolin-3-yl)- acetamide D 106

358.4 2-(4-Oxo-4H- quinazolin-3-yl)-N- (3-sulfamoyl- phenyl)- acetamideD 107

386.4 N-(3- Dimethylsulfamoyl- phenyl)-2-(4-oxo- 4H-quinazolin-3-yl)-acetamide D 108

402.4 4-Oxo-3-[(4- sulfamoyl- phenylcarbamoyl)- methyl]-3,4-dihydro-phthalazine-1- carboxylic acid D 109

372.4 3-(4-Oxo-3,4- dihydro-quinazolin- 2-yl)-N-(4- sulfamoyl-phenyl)-propionamide D 110

403.4 2-(7-Nitro-4-oxo- 4H-quinazolin-3- yl)-N-(4-sulfamoyl-phenyl)-acetamide D 111

416.4 (4-Oxo-4H- quinazolin-3-yl)- acetic acid (4- sulfamoyl-phenylcarbamoyl)- methyl ester D 112

400.5 4-Dimethylsulfamoyl- N-methyl-N-(4-oxo- 3,4-dihydro- quinazolin-2-ylmethyl)-benzamide D 113

413.5 2-(4-Oxo-4H- benzo[d][1,2,3]triazin- 3-yl)-N-[4- (pyrrolidine-1-sulfonyl)- phenyl]-acetamide D 114

443.5 2-[Ethyl-(4-oxo- 3,4-dihydro- quinazolin-2- ylmethyl)-arnino]-N-[2-(4-sulfamoyl- phenyl)-ethyl]- acetamide D 115

443.5 N-(4- Dimethylsulfamoyl- phenyl)-2-[ethyl- (4-oxo-3,4-dihydro-quinazolin-2- ylmethyl)-amino]- acetamide D 116

430.4 3-(4-Oxo-3,4- dihydro-quinazolin- 2-yl)-propionic acid(4-sulfamoyl- phenylcarbamoyl)- methyl ester D 117

477.5 (4-Oxo-4H- quinazolin-3- yl)-acetic acid 4-[(3,4-dimethyl-benzenesulfonyl)- methyl-amino]- phenyl ester D 118

503.6 2-[(6,7-Dimethoxy- 4-oxo-3,4-dihydro- quinazolin-2-ylmethyl)-ethyl- amino]-N-[2-(4- sulfamoyl-phenyl)- ethyl]-acetamide D119

358.4 2-(2-Oxo-2H- quinoxalin-1-yl)- N-(4-sulfamoyl- phenyl)-acetamide D120

475.5 2-[(6,7-Dimethoxy- 4-oxo-3,4-dihydro- quinazolin-2-ylmethyl)-ethyl- amino]-N-(4- sulfamoyl-phenyl)- acetamide D 121

412.5 2-(4-Oxo-4H- quinazolin-3-yl)- N-[3-(pyrrolidine-1-sulfonyl)-phenyl]- acetamide D 122

448.5 N-[3-(Methyl- phenyl-sulfamoyl)- phenyl]-2-(4-oxo-4H-quinazolin-3- yl)-acetamide D 123

457.5 2-(7-Nitro- 4-oxo-4H- quinazolin-3- yl)-N-[4- (pyrrolidine-1-sulfonyl)-phenyl]- acetamide D 124

470.5 (4-Oxo-4H- quinazolin-3- yl)-acetic acid [4- (pyrrolidine-1-sulfonyl)- phenylcarbamoyl]- methyl ester D 125

474.5 (4-Oxo-4H- quinazolin-3-yl)- acetic acid (5- dimethylsulfamoyl-2-methoxy- phenylcarbamoyl)- methyl ester D 126

461.5 N-(5- Dimethylsulfamoyl- 2-methoxy-phenyl)- 2-(7-nitro-4-oxo-4H-quinazolin-3- yl)-acetamide D 127

359.4 4-[2-(4-Oxo-4H- quinazolin-3-yl)- acetylamino]- benzenesulfonicacid D 128

401.4 (4-Oxo-4H- quinazolin-3- yl)-acetic acid 3- dimethylsulfamoyl-benzyl ester D 129

434.9 2- (6-Chloro- 4-oxo-4H- quinazolin-3- yl)-N-(5- dimethylsulfamoyl-2-methyl-phenyl)- acetamide D 130

386.4 2-(2-Oxo-2H- quinoxalin-1-yl)- N-[2-(4-sulfamoyl- phenyl)-ethyl]-acetamide D 131

359.4 2-(4-Oxo-4H- benzo[d][1,2,3]triazin- 3-yl)-N-(3-sulfamoyl-phenyl)- acetamide D 132

414.5 N-Methyl-3-(4- oxo-3,4-dihydro- quinazolin-2-yl)- N-[1-(4-sulfamoyl-phenyl)- ethyl]-propionamide D 133

457.6 N-(4- Isopropylsulfamoyl- phenyl)-2-[methyl- (4-oxo-3,4-dihydro-quinazolin-2- ylmethyl)-amino]- propionamide D 134

471.5 2-(6-Nitro-4- oxo-4H-quinazolin- 3-yl)-N-[4- (piperidine-1-sulfonyl)-phenyl]- acetamide D 135

473.5 N-[4-(Morpholine- 4-sulfonyl)-phenyl]- 2-(6-nitro-4-oxo-4H-quinazolin-3- yl)-acetamide D 136

388.4 N-(4-Methoxy-3- sulfamoyl-phenyl)- 2-(4-oxo-4H- quinazolin-3-yl)-acetamide D 137

476.6 3-(4-Oxo-3-p-tolyl- 3,4-dihydro- quinazolin-2-yl)- N-(4-sulfamoyl-benzyl)- propionamide D 138

475.0 N-[4-(Azepane-1- sulfonyl)-phenyl]- 2-(6-chloro-4-oxo-4H-quinazolin-3- yl)-acetamide D 139

458.5 N-[4-(Azepane-1- sulfonyl)-phenyl]- 2-(7-fluoro-4-oxo-4H-quinazolin- 3-yl)-acetamide D

Example 2 Determining Selectivity or Specificity of Compounds of thePresent Invention

Those compounds with activity against dengue-2 at effectiveconcentrations of less than 10 μM as identified in Example 1 above weretested for activity against each serotype of dengue in a viral yieldassay to generate EC50 values (Table 2). Select compounds were alsotested for more broad spectrum activity against other members of theFlaviviridae family including Modoc, which is a murine flavivirus, aswell as Bovine Viral Diarrhea Virus (BVDV), which is a Pestivirus. Sincedengue virus is able to replicate in multiple cell lines and to ensurethat the activity seen in vero cells is consistent, select compoundswere also tested for their effective concentration in a viral yieldassay against dengue-2 in C6/36 mosquito cells.

TABLE 2 Select Compounds activities against Den-1, Den-2, Den-3, Den-4,Modoc and BVDV. Com- EC50/90 Specificity pound MW Structure (μM) Yield(μM) 1 414.48 MW

4.3/10.7 Den-1 0.3/5.2 Den-2 1.0/8.2 Den-3 1.7/4.1 Den-4 1.2 Modoc >50BVDV 0.3 C6/36 2 328.43 MW

7.6/8.9 Den-1 0.3/4.1 Den-2 5.5/13.3 Den-3 2.1/12.5 Den-4 6.2 Modoc >50BVDV 0.7 C6/36 3 415.47 MW

1.3/5.1 Den-1 4.0/10.0 Den-2 1.0/12.1 Den-3 2.3/10.0 Den-4 3.1 Modoc >50BVDV 4 448.50 MW

7.8/8.9 Den-2 13.5/22.6 Den-3 4.3/22.7 Den-4 >25 Modoc >50 BVDV 6 448.93MW

1.8/43.2 Den-2 64 BVDV 7 432.42 MW

4.6/>50 Den-2 >50 BVDV 8 430.48 MW

2.8/25.6 Den-2 >50 BVDV 9 465.52 MW

1.3/7.3 Den-2 >50 BVDV 10 448.5 MW

1.25/>50 Den-2 11 448.5 MW

4.9/>50 Den-2 12 473.51 MW

>25/>25 Den-1 11.1/>25 Den-2 2.9/>25 Den-3 5.2/14.6 Den-4 >25 Modoc >50BVDV 13 478.53 MW

7.1/8.5 Den-1 2.4/10.3 Den-2 4.1/9.4 Den-3 1.9/9.2 Den-4 >25 Modoc 47.7BVDV 2.1 C6/36 14 328.43 MW

>25/>25 Den-1 3.6/27.5 Den-2 4.2/14.9 Den-3 4.8/11.1 Den-4 >25 Modoc >50BVDV 15 342.46 MW

2.4/>100 Den-2 1.2/37.2 Den-4 10.1 Modoc >50 BVDV 16 376.47 MW

Den-1 5.8/10.3 Den-2 Den-3 2.1/6.4 Den-4 3.6 Modoc >50 BVDV 17

Den-1 5.1/12.1 Den-2 Den-3 Den-4 20 449.94 MW

Den-1 1.3/20 Den-2 Den-3 Den-4 21 342.46 MW

Den-1 14.2/24.1 Den-2 Den-3 8.2/12.0 Den-4 6 Modoc >50 BVDV

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All references cited herein are herein incorporated by reference intheir entirety for all purposes.

The invention has been described in terms of preferred embodimentsthereof, but is more broadly applicable as will be understood by thoseskilled in the art. The scope of the invention is only limited by thefollowing claims.

What is claimed is:
 1. A pharmaceutical composition comprising apharmaceutically acceptable carrier and a compound selected from thegroup consisting ofN-(4-Diethylsulfamoyl-phenyl)-2-(4-oxo-4H-quinazolin-3-yl)-acetamide;N-(5-Diethylsulfamoyl-2-methoxy-phenyl)-isobutyramide;N-(4-Diethylsulfamoyl-phenyl)-2-(4-oxo-4H-benzo[d][1,2,3]triazin-3-yl)-acetamide;N-(4-Diethylsulfamoyl-phenyl)-2-(1,3-dimethyl-2,6-dioxo-1,2,3,6-tetrahydro-purin-7-yl)-acetamide;N-(4-Diethylsulfamoyl-phenyl)-2-(4-oxo-4H-thieno[2,3-d]pyrimidin-3-yl)-acetamide;N-(4-Diethylsulfamoyl-phenyl)-2-(1,4-dioxo-3,4-dihydro-1H-phthalazin-2-yl)-acetamide;N-(4-Diethylsulfamoyl-phenyl)-2-(1,3-dioxo-1H,3H-benzo[de]isoquinolin-2-yl)-acetamide;N-(4-Diethylsulfamoyl-phenyl)-2-(5,6-dimethyl-4-oxo-4H-thieno[2,3-d]pyrimidin-3-yl)-acetamide;N-(4-Diethylsulfamoyl-phenyl)-2-(6-ethyl-4-oxo-4H-thieno[2,3-d]pyrimidin-3-yl)-acetamide;N-(5-Diethylsulfamoyl-2-methyl-phenyl)-2-(7-nitro-4-oxo-4H-quinazolin-3-yl)-acetamide;N-(5-Diethylsulfamoyl-2-methoxy-phenyl)-2-(1,3-dimethyl-2,6-dioxo-1,2,3,6-tetrahydro-purin-7-yl)-acetamide;N-(5-Diethylsulfamoyl-2-methoxy-phenyl)-butyramide;N-(5-Diethylsulfamoyl-2-methoxy-phenyl)-3-methyl-butyramide;N-(5-Diethylsulfamoyl-2-methoxy-phenyl)-2-phenyl-acetamide;1-[2-(4-oxo-4H-quinazolin-3-yl)-acetyl]-2,3-dihydro-1H-indole-5-sulfonicacid diethylamide;N-(5-Diethylsulfamoyl-2-methoxy-phenyl)-2-(4-oxo-4H-benzo[d][1,2,3]triazin-3-yl)-acetamide;Pentanoic acid (5-diethylsulfamoyl-2-methoxy-phenyl)-amide;N-(4-Diethylsulfamoyl-phenyl)-2-(4-oxo-4H-thieno[3,2-d]pyrimidin-3-yl)-acetamide;2-(7-Chloro-4-oxo-4H-quinazolin-3-yl)-N-(4-diethylsulfamoyl-phenyl)-acetamide;N-(4-Diethylsulfamoyl-phenyl)-2-(4-oxo-4H-benzo[4,5]furo[3,2-d]pyrimidin-3-yl)-acetamide;N-(5-Diethylsulfamoyl-2-isopropoxy-phenyl)-3-(4-oxo-4H-benzo[d][1,2,3]triazin-3-yl)-propionamide;N-(4-Diethylsulfamoyl-phenyl)-2-(8-methyl-4-oxo-4H-quinazolin-3-yl)-acetamide;N-(5-Diethylsulfamoyl-2-methoxy-phenyl)-3-(4-oxo-4H-benzo[d][1,2,3]triazin-3-yl)-propionamide;andN-(4-Diethylsulfamoyl-phenyl)-2-[ethyl-(4-oxo-3,4-dihydro-quinazolin-2-ylmethyl)-amino]-acetamide;wherein said pharmaceutical composition is suitable for administrationin a human or animal and wherein said pharmaceutically acceptablecarrier is selected form the group consisting of, dispersion aids,suspension aids, surface active agents, isotonic agents, thickeningagents, emulsifying agents, preservatives, solid binders and lubricants.2. The composition of claim 1, wherein the compound isN-(4-Diethylsulfamoyl-phenyl)-2-(4-oxo-4H-quinazolin-3-yl)-acetamide. 3.The pharmaceutical composition according to claim 1, wherein saidpharmaceutical composition is suitable for administration in a human oranimal, wherein said administration is selected from the groupconsisting of: oral administration, rectal administration, parenteraladministration, intravaginal administration, intraperitonealadministration and administration by inhalation.
 4. The pharmaceuticalcomposition according to claim 1, wherein said pharmaceuticallyacceptable carrier is solid.
 5. The pharmaceutical composition accordingto claim 1, wherein said pharmaceutically acceptable carrier is liquid.6. The pharmaceutical composition according to claim 3, wherein saidparenteral administration is selected from the group consisting of:intramuscular injection, subcutaneous injection and intravenousinfusion.